Blanchet G B, Russell J B, Fincher C R, Portmann M
E.I. DuPont de Nemours, Wilmington, Delaware.
Fertil Steril. 1992 Jun;57(6):1337-41. doi: 10.1016/s0015-0282(16)55097-3.
To introduce the use of excimer lasers for penetration of the zona pellucida for micromanipulation purposes.
Cryopreserved two-cell mouse embryos were thawed and exposed to the 248-nm line of a krypton fluoride excimer laser (Lambda Physik EMG 202, Goettingen, Germany) creating a 2 to 4-micron opening in the zona pellucida.
The Laser Ablation Laboratory at DuPont and the in Vitro Fertilization Laboratory at The Medical Center.
The embryos were exposed in either phosphate-buffered solution (PBS) or modified human tubal fluid (HTF) with the laser power varying from 1 to 2 J/cm2 and cultured in Ham's F-10 medium (GIBCO, Grand Island, NY) with 0.4% bovine serum albumin.
The outcome of each experiment was measured by blastocyst formation of laser-exposed embryos as compared with a set of unexposed control embryos handled in a similar fashion.
Successful laser penetration of the zona pellucida was achieved using the 248-nm line of a krypton fluoride excimer laser. A higher blastocyst formation was found for embryos exposed in PBS. The higher optical absorption of the modified HTF partially inhibited embryo development. The blastocyst statistics increased 2.5-fold times by reducing the exposure of the embryos to ablation by-products.
The use of a krypton fluoride excimer laser was introduced as a new method to open the zona pellucida of two-cell mouse embryos without interrupting blastocyst formation.
介绍准分子激光用于穿透透明带以进行显微操作的用途。
将冷冻保存的二细胞小鼠胚胎解冻，并用氟化氪准分子激光（德国哥廷根Lambda Physik EMG 202）的248纳米谱线照射，在透明带上制造一个2至4微米的开口。
杜邦公司的激光消融实验室和医疗中心的体外受精实验室。
将胚胎置于磷酸盐缓冲液（PBS）或改良的人输卵管液（HTF）中，激光功率在1至2 J/cm²之间变化，并在含有0.4%牛血清白蛋白的哈姆F-10培养基（纽约州大岛市GIBCO公司）中培养。
通过将激光照射的胚胎与一组以类似方式处理的未照射对照胚胎的囊胚形成情况来衡量每个实验的结果。
使用氟化氪准分子激光的248纳米谱线成功实现了透明带的激光穿透。发现置于PBS中的胚胎囊胚形成率更高。改良HTF的较高光吸收部分抑制了胚胎发育。通过减少胚胎与消融副产物的接触，囊胚统计数据增加了2.5倍。
引入了使用氟化氪准分子激光作为一种新方法来打开二细胞小鼠胚胎的透明带，而不影响囊胚形成。
